• Journal of Life Science
• /
• v.21 no.7
• /
• pp.953-960
• /
• 2011

Necrosis is characterized by the cell membrane rupture and release of the cellular contents, including high-mobility group box 1 protein (HMGB1), into the extracellular microenvironment. HMGB1 acts as a transcriptional regulator in nuclei, but exerts a pro-inflammatory and tumor-promoting cytokine activity when released into the extracellular space. Its overexpression is associated with tumor progression and chemoresistance. Thus, HMGB1 acts as a clinically important molecule in tumor biology. In this study, we examined whether HMGB1 affects cell death induced by anti-cancer drugs. Here we show that HMGB1 prevented cisplatin (alkylating agent)-induced apoptosis and switched the cell fate to necrosis in MCF-7, MDA-MB231, and MDA-MB361 cells. Similar apoptosis-to-necrosis switch effects of HMGB1 were observed in cells treated with 4-HC, another alkylating agent. In contrast, HMGB1 did not exert any significant effects on docetaxel (DOC)-induced apoptosis in MCF-7 cells. We also show that cisplatin-induced apoptosis was switched to necrosis in MCF-7 multicellular tumor spheroids (MTS) that were cultured for 8 days and had necrotic cores, but DOC-induced apoptosis was prevented without the apoptosis-to-necrosis switch. Finally, the levels of RAGE, a receptor of HMGB1, were increased with extended culture of MTS. These findings demonstrate that HMGB1 switches alkylating agent-induced apoptosis to necrosis, suggesting that the strategy to prevent necrosis occurring as an undesirable action of alkylating agent-based chemotherapy should be delineated to improve the efficacy of chemotherapy for cancer.

• Journal of Life Science
• /
• v.21 no.7
• /
• pp.1032-1038
• /
• 2011

Recently identified Ciona intestinalis voltage sensor-containing phosphatase (Ci-VSP) consists of an ion channel-like transmembrane domain (VSD) and a phosphatase-like domain. Ci-VSP senses the change of membrane potential by its VSD and works as a phosphoinositide phosphatase by its phosphatase domain. In this study, we present the construction of His-tagged phosphatase-like domain of Ci-VSP, its recombinant expression and purification, and its enzymatic activity behavior in order to examine the biochemical behavior of phosphatase domain of Ci-VSP without interference. We found that Ci-VSP(248-576)-His can be eluted with an elution buffer containing 25 mM NaCl and 100 mM imidazole during His-tag purification. In addition, we found the proper measurement condition for kinetics study of Ci-VSP(248-576)-His against p-nitrophenyl phosphate (pNPP). We measured the kinetic constant of Ci-VSP(248-576)-His at $37^{\circ}C$, pH 5.0 or 5.5, under 30 min of reaction time, and less than $2.0\;{\mu}g$ of protein amount. With these conditions, we acquired that Ci-VSP(248-576)-His has $K_m$ of $354{\pm}0.143\;{\mu}M$, $V_{max}$ of $0.0607{\pm}0.0137\;{\mu}mol$/min/mg and $k_{cat}$ of $0.359{\pm}0.009751\;min^{-1}$ for pNPP dephosphorylation. Therefore, we produced a pure form of Ci-VSP(248-576)-His, and this showed a higher activity against pNPP. This purified protein will provide the road to a structural investigation on an interesting protein, Ci-VSP.

• Journal of Life Science
• /
• v.27 no.10
• /
• pp.1199-1206
• /
• 2017

The lymphotoxin ${\beta}$ receptor ($LT{\beta}R$), a member of the tumor necrosis factor receptor family, plays an important role in lymphoid tissue's architecture and organogenesis. We found that $LT{\beta}R$ stimulation induced changes in stress fibers (SFs) in fibroblastic reticular cells (FRCs). MLCK and ROCK play critical roles in the regulation of SF formation in cells. The present study was performed to investigate the antifibrotic effects on SF regulation of $LT{\beta}R$ signaling, with a focus on MLCK inhibition. The effect of $LT{\beta}R$ on the SF change was analyzed using immunoblot and fluorescence assays and agonistic $anti-LT{\beta}R$ antibody-treated FRCs. In addition, we checked the level of Rho-guanosine diphosphate (GDP)/guanosine triphosphate (GTP) exchange activity with FRC lysate. Phospho-ezrin proteins acting as membrane-cytoskeleton linkers completely de-phosphorylated in agonistic $anti-LT{\beta}R$ antibody-treated FRCs. The actin bundles rearranged into SFs, where phospho-myosin light chain (p-MLC) co-localized in FRCs. ML7-treated FRCs completely blocked SFs and showed retraction and shrinkage processes comparable to those observed in agonistic $anti-LT{\beta}R$ antibody-treated cells. Inhibition of ROCK activity induced changes in the actin cytoskeleton organization; however, some SFs remained in the cells, while they were completely disrupted by MLCK inhibition with ML7. We showed that the phosphorylation of MLC was completely abolished with $LT{\beta}R$ stimulation in FRCs. When $LT{\beta}R$ was stimulated with the agonistic $anti-LT{\beta}R$ antibody, the Rho-GDP/GTP exchange activity was reduced, however, the activity was not completely abolished. Collectively, the results illustrated that MLCK was potently responsible for the SF regulation triggered via $LT{\beta}R$ signaling in FRCs.

• Korean Journal of Food Science and Technology
• /
• v.34 no.2
• /
• pp.255-262
• /
• 2002

This experiment was performed to improve the stability of Lactobacillus fermentum YL-3 as a poultry probiotics. The culture conditions that improve acid tolerance of L. fermentum YL-3 were investigated by changing several factors such as medium composition, temperature, anaerobic incubation and culture time. Also, L. fermentum YL-3 was encapsulated with alginate, calcium chloride and chitosan. The stable culture conditions of L. fermentum YL-3 were obtained in anaerobic incubation using MRS media without tween 80 for 20 hour at $42^{\circ}C$. The capsule after treatment with 1% chitosan was formed close membrane by a bridge bond. Immobilization of L. fermentum YL-3 in capsule was observed by confocal laser scanning microscopy, and cell viability was $2.0{\times}10^9\;CFU/g$ above the average. L. fermentum YL-3 capsule after acid treated at pH 2.0 for 3 hour survived about 40%, but those encapsulated with 1% chitosan survived about 65%. Survival rate of capsule stored at room temperature decreased about $2{\sim}3$ log cycle during 3 weeks, but viability of capsule stored at $4^{\circ}C$ during 3 weeks maintained almost $10^8\;CFU/g$ levels.

• Tuberculosis and Respiratory Diseases
• /
• v.48 no.4
• /
• pp.487-499
• /
• 2000

Background : Acute lung injury is an hypoxic respiratory failure resulting from damage to the alveolar-capillary membrane, which can be developed by a variety of systemic inflammatory diseases. In this study the therapeutic effects of intra-tracheal pulmonary surfactant instillation was evaluated in the intratracheal endotoxin induced acute lung injury model of a rat. Methods : Twenty Sprague-Dawley rats were divided into 4 groups, and normal saline (2 ml/kg, for group 1) or LPS (5 mg/kg, for group 2, 3, and 4) was instilled into the trachea respectively. Either normal saline (2 ml/kg, for group 1 & 2, 30 min later) or bovine surfactant (15 mg/kg, 30 min later for group 3, 5 hr later for group 5) was instilled into the trachea. The therapeutic effect of intratracheal surfactant therapy was evaluated with one chamber body plethysmography (respiratory frequency, tidal volume and enhanced pause), ABGA, BAL fluid analysis (cell count with differential, protein concentration) and pathologic examination of the lung. Results : Intratracheal endotoxin instillation increased the respiration rate decreased the tidal volume and int creased the Penh in all group of rats. Intratracheal instillation of surfactant decreased Penh, increased arterial oxygen tension, decreased protein concentration of BAL fluid and decreased lung inflammation at both times of administration (30 minute and 5 hour after endotoxin instillation). Conclusion : Intratracheal instillation of surfactant can be a beneficial therapeutic modality as discovered in the acute lung injury model of rats induced by intratracheal LPS intillation. It deserves to be evaluated for treatment of human acute lung injury.

• Radiation Oncology Journal
• /
• v.22 no.2
• /
• pp.145-154
• /
• 2004

Purpose : To examine whether a synthetic bile acid derivatives (HS-1200) sensitizes the radiation-induced apoptosis in human breast cancer cells (MCF-7) and to investigate the underlying mechanism. Materials and Methods : Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37$^{\circ}C$ with 5$\%$ CO$_{2}$ in air atmosphere. After removal of HS-1200, cells were irradiated with 2$\~$8 Gy X-ray, and then cultured Ii drug-free media for 24-96 hours. The effect of radiation on the clonogenicity of MCF-7 cells was determined with clonogenic cell survival assay with 16$\mu$M of HS-1200. The induction of apoptosis was determined using agarose gel electrophoresis and Hoechst staining. The expression level of apoptosis-related molecules, such as PARP, Bax, Bcl-2, Bak and AIF, were assayed by Western blotting analysis with 40$\mu$M of HS-1200 combined with 8 Gy irradiation. To examine the cellular location of cytochrome c, bax and AIF immunofluorescent stainings were undertaken. Results : Treatment of MCF-7 cells with 40$\mu$M of HS-1200 combined with 8 Gy irradiation showed several changes associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200 combined with 8 Gy irradiation treatment also enhanced production of PARP cleavage products and increased Bax/Bcl-2 ratio by Western blotting. Loss of mitochondrial membrane potential ($\Delta$$\psi$$_{m}$) and increased cytochrome c staining indicated that cytochrome c had been released from the mitochondria in HS-1200 treated cells. Conclusion : We demonstrated that combination treatment with a synthetic chenodeoxycholic acid derivative HS-1200 and irradiation enhanced radiation-induced apoptosis of human breast cancer cells (MCF-7). We suggest that the increased Bax/Bcl-2 ratio In HS-1200 co-treatment group underlies the increased radio sensitivity of MCF-7 cells. Further futures studies are remained elusive.

• Proceedings of the KOR-BRONCHOESO Conference
• /
• 1979.05a
• /
• pp.3.2-3
• /
• 1979

Bone conduction loss is one of the most common complication in chronic otitis media, and is mostly high frequency loss. Of 233 tympanomastoidectomy ears, 187 ears were considered eligible for this study. A histopathological change was examined in the natural otitis media of guinea pigs. It is our intention to analyze the pattern of bone conduction loss in chronic otitis media, and to correlate this findings with clinical and pathological changes in human and animal otitis media. l) In unilateral cases, a significant difference in bone conduction threshold was observed between normal and diseased ears, and between each frequency with significant interaction between 2KHz and 4KHz (p 0.01). 2) Using one way analysis of variance, mean bone conduction was compared with the duration of disease. We observed a significant difference (p 0.05) between each group of duration, except between 11-15 and 15-20 years group. 3) A comparison of bone conduction between stapes loss group and intact stapes group revealed significant t ratio (p 0.01) at each frequency. The effect of stapes loss on each frequency was evaluated, using one way analysis of variance. there were significant difference(p 0.05) between 250Hz and 500Hz. and between 2KHz and 4KHz. 4) A comparison of bone conduction between round window obliteration and nonobliteration group revealed significant t ratio (p 0.01) at each frequency. Using one way analysis of variance. the effect of round window obliteration was evaluated in each frequency. We observed significant difference (p 0.05) between 250Hz and 500Hz. and between 2KHz and 4KHz. 5) A comparison of bone conduction between cholesteatoma and non -cholesteatoma group revealed significant t ratio (p 0.01) only in 2KHz and 4KHz. No significant differency was observed in mean bone conduction. 6) In a histopathological study of natural otitis media in guinea pig, we observed inflammatory infiltration of the round window membrane, serofibrinous precipitate in the scala tympani, and degeneration of the organ of Corti most significant near the basal turn. These changes would explain high tone bone conduction loss in the process of chronic otitis media.

• Proceedings of the KOR-BRONCHOESO Conference
• /
• 1979.05a
• /
• pp.4.4-5
• /
• 1979

The degree of hearing impairment of chronic otitis media will indicate the nature and severity of middle ear pathology especially condition of ossicular chain, size of ear drum perforation and location of granulation tissue in the middle ear cavity. The subjects were 189 ears of tympanoplasty for chronic otitis media and divided into four groups as follows: Normal ossicular chain with only ear drum perforation (group I), normal ossicular chain with granulation tissue only around the ossicles regardless of any other region (group II), ossicular ankylosis or fixation of handle of malleus to promontory with or without granulation tissue around the ossicle (group III) and ossicular interruption by partial or complete destruction(groupf IV). The results were concluded as follows: 1) The average hearing threshold of chronic otitis media was 44.6 dB and hearing threshold was closely related to the condition of ossicular chain. Hearing threshold became greater in order of normal ossicular chain, ankylosis and interruption. 2) The average hearing threshold of ossicular interrupted group was 49.1 dB and it was greater in the cases of total destruction than that of partial destruction. 3) The hearing loss in the cases of normal ossicular chain with only tympanic membrane perforation was within 45 dB and this level was presumed to represent normal ossicular function. The degree of hearing loss was in proportion to the size of ear drum perforation and when over 45 dB, other middle ear pathology was suggested. 4) In the cases of small ear drum perforation with normal ossicular function, the hearing threshold was within 30 dB. 5) In the type of audiogram, flat type was 30.2% and ascending type 35.4%. Descending type was more frequent in the cases of normal ossicular mobility with granulation tissue around the ossicle and flat type was observed frequently in the cases of ossicular ankylosis. 6) Carhart's notch was seen in 14 cases (7.4%) and observed mainly in ossicular ankylosis. 7) There was no relation between hearing threshold and histopathological type of granulation tissue in chronic otitis media. However the degree of hearing impairment was related to the location of granulation tissue in the middle ear cavity. 8) Authors recognized the granulation tissue compensated the function of interrupted ossicular chain.

• Journal of dental hygiene science
• /
• v.13 no.4
• /
• pp.386-392
• /
• 2013

The purpose of this study was to evaluate the factors related to xerostomia in patients with systemic diseases, and the results were analyzed through t-test, ANOVA, and multiple linear regression analysis. There were 62 respondents (56.9%) who reported dry mouth and the season of the most severe dry mouth was reported to be the highest during winter. According to the analysis made in the relationship of xerostomia with the awareness of general health and health behavior, the group that understood their own health poorly, had more than two kinds of diseases, and had been administered more than two kinds of daily medication showed higher xerostomia. Also, those who experienced desperation, had high dryness in other parts of the body, and heavy snoring were more aware of xerostomia (p<0.05). According to the analysis made in the relationship between xerostomia and awareness of oral health state, the group that understood their oral health poorly and had pain in the oral mucous membrane and halitosis was more aware of xerostomia (p<0.05). According to the analysis made in the relationship between quality of life and xerostomia, the group that showed high level of problem in daily living and stress and were self-conscious and felt tense due to halitosis reported more xerostomia (p<0.05). Finally, xerostomia was most correlated with consciousness of the eyes of others due to halitosis followed by the number of transferred systemic diseases and stress level. Based on such results, xerostomia was understood to be an obstacle in maintaining favorable social life and health. Since xerostomia was shown to be related to the number of transferred systemic diseases, the dental professions are asked to further improve their understanding not only on oral health but also systemic diseases. Therefore, these efforts are expected to prevent xerostomia and reduce various complications caused by xerostomia.

• Clean Technology
• /
• v.10 no.2
• /
• pp.73-87
• /
• 2004

In most of industrial fields, cleaning is employed for removing soils on their products or parts. Halogenated cleaning agents such as CFC-113, 1,1,1-TCE(1,1,1-trichloroethane), MC(methylene chloride) and TCE (trichloroethylene) have been used as cleaning ones in most of companies in the world since their excellent performance of cleaning ability and good material compatibility. However, CFC-113 and 1,1,1-TCE which are ozone destruction substances are not used any more in the advanced countries because of the which are ozone destruction substances are not used any more in the advanced countries because of the Montreal protocol. MC and TCE are now used restrictively at small part of industrial fields in most of countries since they are known to be hazardous or carcinogenic materials. Thus, it is indispensible that the alternative cleaning agents which are environmental-friendly and safe, and show good cleaning ability should be developed or utilized for replacement of the halogenated cleaning agents. Aqueous/semi-aqueous cleaning agents are evaluated to be promising alternative ones among various alternatives in environmental and economical view point. In this study, commercially available 12 aqueous and 6 semi-aqueous cleaning agents were selected and their physical properties, cleaning abilities, rinsing abilities and recycling of contaminated rinse water were measured and analyzed. Aqueous cleaning agents with higher wetting index showed better cleaning ability compared with those with lower wetting index. However wetting index did not have any correlation with cleaning ability in semi-aqueous cleaning agents. It was observed that soil concentration in aqueous and semi-aqueous cleaning agents should be maintained below the certain concentrations which depend on types of clearing agents. More than 70% soils in contaminated rinse water by some of aqueous and semi-aqueous clearing agents could be separated by simple settling method. This means that some cleaning agents with high oil-water separation efficiency will be effiective for recycling oil-contaminated rinse water. It was found that contaminated rinse water with aqueous agents was purified easiy by ultrafiltration method with PAN membrane of 30 kDa.